Figure 5
LX2343 ameliorated tau pathology involving GSK-3β inhibition. (A–D) Western blot and quantification results demonstrated that LX2343 reduced tau phosphorylation at sites of serine 396/199 and threonine 231 in SH-SY5Y and primary neuronal cells (one-way ANOVA, Dunnett's multiple comparison test, *P<0.05, **P<0.01 versus STZ. #P<0.05, ##P<0.01 versus DMSO). (E–H) Western blot and quantification results demonstrated that LX2343 reduced GSK-3β phosphorylation at tyrosine 216 and increased GSK-3β phosphorylation at serine 9 in SH-SY5Y and primary neuronal cells (one-way ANOVA, Dunnett's multiple comparison test. *P<0.05, **P<0.01 versus STZ. ##P<0.01 versus DMSO). (I) LX2343 dose-dependently inhibited GSK-3β activity in vitro (one-way ANOVA, Dunnett's multiple comparison test. *P<0.05, **P<0.01 versus DMSO; SB216763: t test. ##P<0.01 versus DMSO). SB216763: GSK-3β inhibitor. (J) LX2343 dose-dependently inhibited GSK-3β in the presence of the indicated concentrations of ATP. In the presence of 10 μmol/L of ATP, the IC50 of LX2343 was 1.37±0.47 μmol/L; in the presence of 30 μmol/L ATP, the IC50 of LX2343 was 1.84±0.07 μmol/L; and in the presence of 100 μmol/L ATP, the IC50 of LX2343 was 1.42±0.09 μmol/L. The LX2343 concentration was expressed in the log10 scale. All values were presented as the mean±SEM (n=3). GAPDH was used as the loading control in the Western blot assays. Data were obtained from three independent experiments.
